help with cryostat cutting

[Brian Scott]

In article <4l5kms$jio at news.cuny.edu>,  <jcoleman at msvax.mssm.edu> wrote:
>In article <4l375j$qt4 at amgen.amgen.com>, none <none> writes:
>>I'm no expert, in fact, I'm not even skilled in this myself.  But
>>just based on what you describe, it sounds like a problem with
>>the smoothness of the blade-falling-action, if you know what I
>>mean.  Is this a manual one, or automatic?  I've only even tried
>>cutting a few times myself, and found it very frustrating, and the
>>problem I encountered most was the one you describe.  I always
>>ascribed it to the speed/motion of the blade across the sample.
>>Perhaps this is just too simplistic, especially since you have
>>years of experience.
>>*** Disclaimer: These are the opinions of the poster not Amgen Inc.***

>Likewise, I am no expert in fact I am most likely to question my choice of
>science as a career when cryostating ...could it be poor sucrose infiltration?
>or perhaps the block itself is too cold.
>
>Just a feeble thought :-)

I usually left the tissue sitting in the sucrose a number of days.  I 
slided brain tissue for 3.5 years in another lab and never had any 
problems.  It might be a mechanical problem although it's not obvious.  
I've always wondered if the fixative had anything to do with it and 
everyone I asked said it wouldn't make a difference.  I just tried 
cutting two pieces of tissue but I used a paraformaldehyde/picric adec 
mixture as a fixative on one instead of my regular.  The one done in the 
regular came out shredded as usual but the other one was perfect.  I'm 
repeating it with both fixatives but longer fixation time to see if it's 
consistant.  I hope it is.

Brian



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Brian Scott            | "In other studies you go as far as others have gone
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